Molecular Pharmacology Laboratory - Honours projects available in 2009
An Honours project undertaken in this lab would be administered by the Discipline of Pharmacology.
- Molecular dynamics of glutamate transporters
Supervisor + contact details:
- Dr Renae Monique Ryan
- Associate Professor Robert Vandenberg
The aim of this project is to develop a molecular model for the mechanism of glutamate transport and through this to identify novel means of pharmacologically manipulating transporter function.
Glutamate transporters play an important role in shaping the dynamics of neurotransmission in the human brain but also have the capacity to create widespread pathological changes. Dysfunction or downregulation of glutamate transporters has been implicated in disease states such as Alzheimer¡¯s disease, motor neuron disease and ischemic brain damage following a stroke. The determination of a crystal structure of the bacterial glutamate transporter GltPh has created great excitement because we now have the tools to begin to understand how these important proteins work. This projects will capitalise on this breakthrough by developing a structural model of how human glutamate transporters work and from this it should be possible to develop novel pharmacological tools to manipulate transporter function.
Using a range of techniques, the molecular basis for transport will be investigated for both the human (EAAT1) and bacterial (Glt Ph) glutamate transporters. Recombinant GltPh protein will be purified and reconstituted into liposomes and 3H-labelled substrate and ion fluxes will be measured. EAAT1 will be expressed in Xenopus laevis oocytes and both 3H-labelled substrate fluxes and two electrode voltage clamp electrophysiology will be used to study function. Site-directed mutagenesis will be used on both transporters to identify functionally important sites and to probe conformational changes upon binding of glutamate and the various coupled ions. Proximity relationships will also be investigated to gain insight into the conformational changes this protein undergoes during substrate translocation.
This information will be invaluable for understanding the role of glutamate and glutamate transporters in normal and disease states. See Boudker, Ryan et al., (2007) Nature 445: 387-393.
- Drug binding sites on glycine transporters
Supervisor + contact details:
- Dr Renae Monique Ryan
- Associate Professor Robert Vandenberg
The aim of this project is develop a structural model for how drugs interact with Glycine Transporters, which will be used in drug design.
Glycine transporters regulate both excitatory and inhibitory neurotransmission in the brain and spinal cord. In the last couple of years it has become apparent that Glycine transport inhibitors may be useful in the treatment of schizophrenia and pain. The determination of the crystal structure of a related bacterial transporter has greatly aided in understanding how these proteins work and also provides tools for understanding how drugs may interact with the transporters.
The Transporter Biology Group in the Discipline of Pharmacology has established that extracellular loops 2 and 4 form part of the binding site for a novel GLYT2 inhibitor, N-arachidonyl-glycine and in this project you will extend this investigation to the roles of amino acid residues of transmembrane domain 3 and 10 of GLYT1 and GLYT2 in forming drug binding sites. This information will provide the basis for the development of new GLYT inhibitors to treat pain. The project will involve site-directed mutagenesis of recombinant Glycine Transporters, expression of the transporters in Xenopos laevis oocytes and electrophysiological analysis of the functional properties of the mutant transporters. There will also be some computer modelling of drug: transporter interactions.
Research in the Transport Biology Group is supported by NHMRC project grants.
